Co-selection of antibiotic and metal resistance

There is growing concern that metal contamination functions as a selective agent in the proliferation of antibiotic resistance. Documented associations between the types and levels of metal contamination and specific patterns of antibiotic resistance suggest that several mechanisms underlie this co-selection process. These co-selection mechanisms include co-resistance (different resistance determinants present on the same genetic element) and cross-resistance (the same genetic determinant responsible for resistance to antibiotics and metals). Indirect but shared regulatory responses to metal and antibiotic exposure such as biofilm induction also represent potential co-selection mechanisms used by prokaryotes. Metal contamination, therefore, represents a long-standing, widespread and recalcitrant selection pressure with both environmental and clinical importance that potentially contributes to the maintenance and spread of antibiotic resistance factors.     

畜禽养殖废物中抗生素和重金属抗性基因的产生机制和控制方法研究进展

动物饲料中常混有抗生素和重金属,导致外排的动物粪便中携带有抗生素和重金属,引发细菌产生耐药性和重金属抗性,继而产生抗生素抗性基因和重金属抗性基因。抗生素和重金属抗性基因污染已成为威胁人类身体健康及破坏生态环境的重大问题。本文从细菌进化的角度,明确了细菌的抗生素和重金属长期进化试验对抗性机制研究的重要性;抗生素抗性基因与重金属抗性基因间存在复杂的协同选择抗性,两者间相互影响,共同决定着细菌环境行为;抗性基因的水平转移增加了细菌在环境中的可变性,可移动遗传元件在抗性基因水平转移中发挥着重要作用。在抗性基因污染控制方面,高级氧化技术具有很好的抗性基因去除效果,尤其是UV/TiO₂氧化技术,能使抗生素抗性基因丰度减少4.7～5.8 log,减少率大于99.99%。其他的控制策略,如抗生素替代品博落回提取物以及噬菌体与抗生素结合使用,对于抗性基因的控制也具有重要意义。     

Co-occurrence of antibiotic,biocide, and heavy metal resistance genes in bacteria from metal and radionuclide contaminated soils at the Savannah River Site

Contaminants such as heavy metals may contribute to the dissemination of antimicrobial resistance (AMR) by enriching resistance gene determinants via co-selection mechanisms. In the present study, a survey was performed on soils collected from four areas at the Savannah River Site (SRS), South Carolina, USA, with varying contaminant profiles: relatively pristine (Upper Three Runs), heavy metals (Ash Basins), radionuclides (Pond B) and heavy metal and radionuclides (Tim’s Branch). Using 16S rRNA gene amplicon sequencing, we explored the structure and diversity of soil bacterial communities. Sites with legacies of metal and/or radionuclide contamination displayed significantly lower bacterial diversity compared to the reference site. Metagenomic analysis indicated that multidrug and vancomycin antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) including those associated with copper, arsenic, iron, nickel and zinc were prominent in all soils including the reference site. However, significant differences were found in the relative abundance and diversity of certain ARGs and MRGs in soils with metal/radionuclide contaminated soils compared to the reference site. Co-occurrence patterns revealed significant ARG/MRG subtypes in predominant soil taxa including Acidobacteriaceae, Bradyrhizobium, Mycobacterium, Streptomyces, Verrumicrobium, Actinomadura and Solirubacterales. Overall, the study emphasizes the potential risk of human activities on the dissemination of AMR in the environment.     

金属离子铁和锌对鲍曼不动杆菌的抗菌作用机制研究进展

由于抗菌药物的开发周期越来越长,远远赶不上细菌耐药的发展速度,临床鲍曼不动杆菌多重耐药与泛耐药现象日益严重。因此,人们越来越关注对抗菌药物以外的抗菌物质的开发,尤其是从生存条件方面来研究抑制耐药菌活性的方法,如金、银、铜等金属离子对鲍曼不动杆菌的作用。本文主要综述铁、锌等金属离子及其螯合物对鲍曼不动杆菌的抗菌作用。铁、锌等金属离子通过与一系列酶的协同作用,调控外排泵或影响生物膜形成及其黏附性等来抑制细菌生长。此外,一些非必需金属如金、银、钯等对鲍曼不动杆菌也具有很强的毒性,有良好的抗菌和降低耐药率的效果,可作为医疗留置器械的抗菌涂层等来预防感染。     

Metal tolerance assisted antibiotic susceptibility profiling in <Emphasis Type="Italic">Comamonas acidovorans</Emphasis>

Metal ions are known selective agents for antibiotic resistance and frequently accumulate in natural environments due to the anthropogenic activities. However, the action of metals that cause the antibiotic resistance is not known for all bacteria. The present work is aimed to investigate the co-selection of metals and antibiotic resistance in Comamonas acidovorans. Tolerance profile of 16 metals revealed that the strain could tolerate high concentrations of toxic metals i.e., Cr (710 ppm), As (380 ppm), Cd (320 ppm), Pb (305 ppm) and Hg (205 ppm). Additionally, metal tolerant phenotypes were subjected to antibiotic resistance profiling; wherein several metal tolerant phenotypes (Cr 1.35-fold; Co-1.33 fold; Mn-1.29 fold) were resistant, while other metal tolerant phenotypes (Mg 1.32-fold; Hg 1.29-fold; Cu 1.28-fold) were susceptible than control phenotype. Metal accumulation may alter the metabolism of C. acidovorans that activates or inactivates the genes responsible for antibiotic resistance, resulting in the resistance and/or susceptibility pattern observed in metal resistant phenotypes.     

Understanding molecular mechanisms for improving phytoremediation of heavy metal-contaminated soils

Heavy metal pollution of soil is a significant environmental problem with a negative potential impact on human health and agriculture. Rhizosphere, as an important interface of soil and plants, plays a significant role in phytoremediation of contaminated soil by heavy metals, in which, microbial populations are known to affect heavy metal mobility and availability to the plant through release of chelating agents, acidification, phosphate solubilization and redox changes, and therefore, have potential to enhance phytoremediation processes. Phytoremediation strategies with appropriate heavy metal-adapted rhizobacteria or mycorrhizas have received more and more attention. In addition, some plants possess a range of potential mechanisms that may be involved in the detoxification of heavy metals, and they manage to survive under metal stresses. High tolerance to heavy metal toxicity could rely either on reduced uptake or increased plant internal sequestration, which is manifested by an interaction between a genotype and its environment.A coordinated network of molecular processes provides plants with multiple metal-detoxifying mechanisms and repair capabilities. The growing application of molecular genetic technologies has led to an increased understanding of mechanisms of heavy metal tolerance/accumulation in plants and, subsequently, many transgenic plants with increased heavy metal resistance, as well as increased uptake of heavy metals, have been developed for the purpose of phytoremediation. This article reviews advantages, possible mechanisms, current status and future direction of phytoremediation for heavy-metal–contaminated soils.     

Bacterial tolerances to metals and antibiotics in metal-contaminated and reference streams

Anthropogenic-derived sources of selection are typically implicated as mechanisms for maintaining antibiotic resistance in the environment. Here we report an additional mechanism for maintaining antibiotic resistance in the environment through bacterial exposure to metals. Using a culture-independent approach, bacteria sampled along a gradient of metal contamination were more tolerant of antibiotics and metals compared to bacteria from a reference site. This evidence supports the hypothesis that metal contamination directly selects for metal tolerant bacteria while co-selecting for antibiotic tolerant bacteria. Additionally, to assess how antibiotic and metal tolerance may be transported through a stream network, we studied antibiotic and metal tolerance patterns over three months in bacteria collected from multiple stream microhabitats including the water column, biofilm, sediment and Corbicula fluminea (Asiatic clam) digestive tracts. Sediment bacteria were the most tolerant to antibiotics and metals, while bacteria from Corbicula were the least tolerant. Differences between microhabitats may be important for identifying reservoirs of resistance and for predicting how these genes are transferred and transported in metal-contaminated streams. Temporal dynamics were not directly correlated to a suite of physicochemical parameters, suggesting that tolerance patterns within microhabitats are linked to a complex interaction of the physicochemical characteristics of the stream.     

Spatial Analysis of Antibiotic Resistance Along Metal Contaminated Streams

The spatial pattern of antibiotic resistance in culturable sediment bacteria from four freshwater streams was examined. Previous research suggests that the prevalence of antibiotic resistance may increase in populations via indirect or coselection from heavy metal contamination. Sample bacteria from each stream were grown in media containing one of four antibiotics—tetracycline, chloramphenicol, kanamycin, and streptomycin—at concentrations greater than the minimum inhibitory concentration, plus a control. Bacteria showed high susceptibilities to the former two antibiotics. We summarized the latter two more prevalent (aminoglycoside) resistance responses and ten metals concentrations per sediment sample, by Principal Components Analysis. Respectively, 63 and 58% of the variability was explained in the first principal component of each variable set. We used these multivariate summary metrics [i.e., first principal component (PC) scores] as input measures for exploring the spatial correlation between antibiotic resistance and metal concentration for each stream sampled. Results show a significant and negative correlation between metals PC scores versus aminoglycoside resistance scores and suggest that selection for metal tolerance among sediment bacteria may influence selection for antibiotic resistance differently in sediments than in the water column. Our most important finding comes from geostatistical cross-variogram analysis, which shows that increasing metal concentration scores are spatially associated with decreasing aminoglycoside resistance scores—a negative correlation, but holds for contaminated streams only. We suspect our field results are influenced by metal bioavailability in the sediments and by a contaminant promoted interaction or “cocktail effect” from complex combinations of pollution mediated selection agents.     

Swarming motility: a multicellular behaviour conferring antimicrobial resistance

Swarming is a type of social motility allowing the migration of highly differentiated bacterial cells. Swarming shares many similarities with biofilm communities, which are notable for their high resistance to antimicrobial agents. We investigate here if the swarming behaviour could also be associated with a widespread antimicrobial resistant phenotype. Challenged with 13 antibiotics from various classes, swarm cells of Pseudomonas aeruginosa , Escherichia coli , Serratia marcescens , Burkholderia thailandensis and Bacillus subtilis showed higher resistance than their planktonic counterparts to all the antibiotics tested, except for the antimicrobial peptides. Using P. aeruginosa as a model, this multiresistant phenotype was shown to be transient and intrinsically linked to the swarming state. Resistance of swarm cells towards other antimicrobial agents, such as triclosan and a heavy metal (arsenite), was also observed. Neither RND-type efflux pumps, including MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM, nor a biofilm-associated resistance mechanism involving periplasmic glucans, appear to account for the resistance of swarm cells. Together with the high resistance of biofilms, these results support the hypothesis that antimicrobial resistance is a general feature of bacterial multicellularity. Swarming motility might thus represent a form of social behaviour useful as a model to investigate biofilm antibiotic resistance.     

Mechanisms of metal resistance in plants: aluminum and heavy metals

Plants have evolved sophisticated mechanisms to deal with toxic levels of metals in the soil. In this paper, an overview of recent progress with regards to understanding fundamental molecular and physiological mechanisms underlying plant resistance to both aluminum (Al) and heavy metals is presented. The discussion of plant Al resistance will focus on recent advances in our understanding of a mechanism based on Al exclusion from the root apex, which is facilitated by Al-activated exudation of organic acid anions. The consideration of heavy metal resistance will focus on research into a metal hyperaccumulating plant species, the Zn/Cd hyperaccumulator, Thlaspi caerulescens, as an example for plant heavy metal research. Based on the specific cases considered in this paper, it appears that quite different strategies are used for Al and heavy metal resistance. For Al, our current understanding of a resistance mechanism based on excluding soil-borne Al from the root apex is presented. For heavy metals, a totally different strategy based on extreme tolerance and metal hyperaccumulation is described for a hyperaccumulator plant species that has evolved on naturally metalliferous soils. The reason these two strategies are the focus of this paper is that, currently, they are the best understood mechanisms of metal resistance in terrestrial plants. However, it is likely that other mechanisms of Al and/or heavy metal resistance are also operating in certain plant species, and there may be common features shared for dealing with Al and heavy resistance. Future research may uncover a number of novel metal resistance mechanisms in plants. Certainly the complex genetics of Al resistance in some crop plant species, such as rice and maize, suggests that a number of presently unidentified mechanisms are part of an overall strategy of metal resistance in crop plants.     

Associations between sensitivity to antibiotics,disinfectants and heavy metals in natural,clinical and laboratory isolates of Escherichia coli

Bacteria in nature often encounter non-antibiotic antibacterials (NAAs), such as disinfectants and heavy metals, and they can evolve resistance via mechanisms that are also involved in antibiotic resistance. Understanding whether susceptibility to different types of antibacterials is non-randomly associated across natural and clinical bacteria is therefore important for predicting the spread of resistance, yet there is no consensus about the extent of such associations or underlying mechanisms. We tested for associations between susceptibility phenotypes of 93 natural and clinical Escherichia coli isolates to various NAAs and antibiotics. Across all compound combinations, we detected a small number of non-random associations, including a trio of positive associations among chloramphenicol, triclosan and benzalkonium chloride. We investigated genetic mechanisms that can explain such associations using genomic information, genetic knockouts and experimental evolution. This revealed some mutations that are selected for by experimental exposure to one compound and confer cross-resistance to other compounds. Surprisingly, these interactions were asymmetric: selection for chloramphenicol resistance conferred cross-resistance to triclosan and benzalkonium chloride, but selection for triclosan resistance did not confer cross-resistance to other compounds. These results identify genetic changes involved in variable cross-resistance across antibiotics and NAAs, potentially contributing to associations in natural and clinical bacteria.     

Understanding cellular responses to toxic agents: a model for mechanism-choice in bacterial metal resistance

Summary Bacterial resistances to metals are heterogeneous in both their genetic and biochemical bases. Metal resistance may be chromosomally-, plasmid- or transposonencoded, and one or more genes may be involved; at the biochemical level at least six different mechanisms are responsible for resistance. Various types of resistance mechanisms can occur singly or in combination and for a particular metal different mechanisms of resistance can occur in the same species. To understand better the diverse responses of bacteria to metal ion challenge we have constructed a qualitative model for the selection of metal resistance in bacteria. How a bacterium becomes resistant to a particular metal depends on the number and location of cellular components sensitive to the specific metal ion. Other important selective factors include the nature of the uptake systems for the metal, the role and interactions of the metal in the normal metabolism of the cell and the availability of plasmid (or transposon) encoded resistance mechanisms. The selection model presented is based on the interaction of these factors and allows predictions to be made about the evolution of metal resistance in bacterial populations. It also allows prediction of the genetic basis and of mechanisms of resistance which are in substantial agreement with those in well-documented populations. The interaction of, and selection for resistance to, toxic substances in addition to metals, such as antibiotics and toxic analogues, involve similar principles to those concerning metals. Potentially, models for selection of resistance to any substance can be derived using this approach.     

Metal and antibiotic resistance in psychrotrophic bacteria associated with the Antarctic sponge Hemigellius pilosus (Kirkpatrick, 1907)

High concentrations of heavy metals have been previously detected in Antarctic sponge tissues, but their effect on the associated bacterial assemblages has been never investigated. Metal tolerance is often linked to antibiotic resistance and can also affect biochemical activities within microbial populations. In the present work, the response to heavy metals and antibiotics, as well as the enzymatic profile, of bacteria associated with the sponge Hemigellius pilosus, was analyzed. Tolerance to mercury, cadmium and zinc (at concentrations between 10 and 10,000 ppm) was tested by the plate diffusion method. Almost all isolates completely tolerated zinc and cadmium up to 1,000 and 2,500 ppm, respectively, whereas complete tolerance to mercury was generally observed at concentrations between 10 and 500 ppm. As bacteria can develop resistance in the growing presence of toxic compounds in the environment, this finding could be related to the concentrations of metals in the sponge tissues. The susceptibility assay to 11 antibiotics revealed that multiple antibiotic resistance was generally exhibited, with gentamicin that inhibited all Antarctic isolates. The comparison of the heavy metal and antibiotic resistance patterns at phylogenetic level revealed some distinctive features, suggesting that the dissemination of heavy metal tolerance and antibiotic resistance may possess great relevance for the population dynamics. Additionally, growth patterns often highly differed among strains in the same species, thus appearing to be more likely strain specific rather than species specific. The enzyme expression by the isolates was not really affected by the heavy metal tolerance they showed, as variation in the enzymatic profiles was observed in strains within the same genus that showed different/similar heavy metal tolerance patterns.     

植物根际促生菌及丛枝菌根真菌协助植物修复重金属污染土壤的机制

植物修复是一种前景广阔的重金属污染土壤的主要修复技术,在微生物的协助下效果更为显著。植物根际促生菌可通过分泌吲哚-3-乙酸(IAA)、产铁载体、固氮溶磷等方式促进植物生长、改善植物重金属耐受性,从而有效提高重金属污染土壤的植物修复效率。菌根真菌是土壤-植物系统中重要的功能菌群之一,可侵染植物根系改变根系形态和矿质营养状况,通过菌丝体吸附重金属,也可产生球囊霉素、有机酸、植物生长素等次生代谢产物改变重金属生物有效性。植物根际促生菌与丛枝菌根真菌可对植物产生协同促生作用,在重金属污染土壤修复中具有一定应用潜力。目前,国内外关于植物根际促生菌和丛枝菌根真菌互作已有大量研究,而二者的相互作用机理仍处于探索阶段。本文综述了近年来国内外植物根际促生菌和丛枝菌根真菌在重金属污染土壤植物修复中的作用机制,并对其研究前景进行展望。     

Potential of 2, 2′-dipyridyl diselane as an adjunct to antibiotics to manage cadmium-induced antibiotic resistance in <Emphasis Type="Italic">Salmonella enterica</Emphasis> serovar Typhi Ty2 strain

One of the reasons for increased antibiotic resistance in Salmonella enterica serovar Typhi Ty2 is the influx of heavy metal ions in the sewage, from where the infection is transmitted. Therefore, curbing these selective agents could be one of the strategies to manage the emergence of multidrug resistance in the pathogen. As observed in our earlier study, the present study also confirmed the links between cadmium accumulation and antibiotic resistance in Salmonella. Therefore, the potential of a chemically-synthesised compound 2, 2′-dipyridyl diselane (DPDS) was explored to combat the metal-induced antibiotic resistance. Its metal chelating and antimicrobial properties were evidenced by fourier transform infrared spectroscopy (FTIR), field emission scanning electron microscopy (FE-SEM), and microbroth dilution method. Owing to these properties of DPDS, further, this compound was evaluated for its potential to be used in combination with conventional antibiotics. The data revealed effective synergism at much lower concentrations of both the agents. Thus, it is indicated from the study that the combination of these two agents at their lower effective doses might reduce the chances of emergence of antibiotic resistance, which can be ascribed to the multi-pronged action of the agents.     

植物对重金属耐性的分子生态机理

植物适应重金属元素胁迫的机制包括阻止和控制重金属的吸收、体内螯合解毒、体内区室化分隔以及代谢平衡等。近年来,随着分子生物学技术在生态学研究中的深入应用,控制这些过程的分子生态机理逐渐被揭示出来。菌根、根系分泌物以及细胞膜是控制重金属进入植物根系细胞的主要生理单元。外生菌根能显著提高寄主植物的重金属耐性,根系分泌物通过改变根际pH、改变金属物质的氧化还原状态和形成络合物等机理减少植物对重金属的吸收。目前,控制菌根和根系分泌物重金属抗性的分子生态机理还不清楚。但细胞膜跨膜转运器已得到深入研究,相关金属离子转运器被鉴定和分离,一些控制基因如铁锌控制运转相关蛋白(ZIP)类、自然抵抗相关巨噬细胞蛋白(Nramp)类、P_1B-type ATPase类基因已被发现和克隆。金属硫蛋白(MTs)、植物螯合素(PCs)、有机酸及氨基酸等是植物体内主要的螯合物质,它们通过螯合作用固定金属离子,降低其生物毒性或改变其移动性。与MTs合成相关的MT-like基因已经被克隆,PCs合成必需的植物螯合素合酶(PCS), 即γ-Glu-Cys二肽转肽酶(γ-ECS) 的编码基因已经被克隆,控制麦根酸合成的氨基酸尼克烟酰胺(NA)在重金属耐性中的作用和分子机理也被揭示出来。ATP 结合转运器(ABC)和阳离子扩散促进器(CDF) 是植物体内两种主要膜转运器,通过它们和其它跨膜方式,重金属被分隔贮藏于液泡内。控制这些蛋白转运器合成的基因也已经被克隆,在植物中的表达证实其与重金属的体内运输和平衡有关。热休克蛋白(HSP)等蛋白类物质的产生是一种重要的体内平衡机制,其分子机理有待进一步研究。重金属耐性植物在这些环节产生了相关响应基因或功能蛋白质,分子克隆和转基因技术又使它们在污染治理上得到了初步的应用。     

The biological cost of mutational antibiotic resistance: any practical conclusions?

A key parameter influencing the rate and trajectory of the evolution of antibiotic resistance is the fitness cost of resistance. Recent studies have demonstrated that antibiotic resistance, whether caused by target alteration or by other mechanisms, generally confers a reduction in fitness expressed as reduced growth, virulence or transmission. These findings imply that resistance might be reversible, provided antibiotic use is reduced. However, several processes act to stabilize resistance, including compensatory evolution where the fitness cost is ameliorated by additional mutation without loss of resistance, the rare occurrence of cost-free resistance mechanisms and genetic linkage or co-selection between the resistance markers and other selected markers. Conceivably we can use this knowledge to rationally choose and design targets and drugs where the costs of resistance are the highest, and where the likelihood of compensation is the lowest.     

解磷微生物修复土壤重金属污染研究进展

土壤重金属污染问题日益严重,具有普遍性、隐蔽性、表聚性、不可逆性等特点,已经成为环境污染治理中的热点、难点问题。解磷微生物能够依靠自身的代谢产物或通过与其他生物的协同作用,将土壤中的难溶性磷转化为可供植物吸收利用的磷,具有多重植物促生长功能和重金属解毒能力,可在重金属毒害水平下,促进植物生长、提高植物抗病能力、克服重金属对植物生长的不利影响,从而增强重金属修复植物的生存竞争力。从解磷微生物的研究现状入手,介绍了解磷微生物对土壤重金属污染的修复能力,综述了解磷微生物对土壤重金属污染修复的作用机制,分析了目前解磷微生物在重金属修复过程中存在的问题,并提出了今后研究的方向,为重金属污染土壤的修复提供了新思路。